Image - Barts and The London logo and link to home page Image - divider Image - divider
  Institute of Cell and Molecular Science
Flow Cytometry Core Facility equipment, images used with permission by BD
  Flow Cytometry Core Facility
  link Home link Instruments available link Flow cytometry link Uses of flow cytometry link Flow sorting link Links link Contacts

FRET based detection of BrdU


FRET can used to measure BrdU incorporation by the use of DNA dyes PI and ToPro3. The incorporation of BrdU is normally analysed in flow cytometry by labelling with a conjugate anti-BrdU mcab and PI to perform cell cycle analysis. This FRET based method relies upon the fact that when DNA is pulsed with BrdU the binding of PI and ToPro3 changes and are brought closer together (<10 nm) allowing FRET to occur resulting in an energy transfer from PI (donor) to ToPro3 (acceptor). Hence on a dot-plot of PI vs ToPro3, the BrdU containing cells have an enhanced ToPro3 signal compared to unpulsed cells. In longer time courses (>12h BrdU pulse) it is possible to track cell generations in a similar manner to that shown in CFSE proliferation assays, with advantage that cell cycle analysis data is collected.







FRET detection of BrdU

FRET Efficiency Ratio Formula


by Gary Warnes. © Queen Mary, University of London 2007
Institute of Cell and Molecular Science, The Blizard Building, Barts and The London School of Medicine and Dentistry, 4 Newark Street, London E1 2AT, UK Tel: +44 (0)20 7882 2483, Fax: +44 (0)20 7882 2200